Neutrophils activated by BJcuL, a C-type lectin isolated from Bothrops jararacussu venom, decrease the invasion potential of neuroblastoma SK-N-SH cells in vitro.

Jessica Ohana Lemes Carneiro-Goetten,Rodrigo Amauri Nogoceke,Andrea Novais Moreno-Amaral,Patricia Maria Stuelp-Campelo,Bruna Santos Rodrigues,Thatyanne Gradowski Do Nascimento,Selene Elifio-Esposito

doi:10.1590/1678-9199-jvatitd-2019-0073

Jessica Ohana Lemes Carneiro-Goetten, Rodrigo Amauri Nogoceke + Show 5 more

Open Access

https://doi.org/10.1590/1678-9199-jvatitd-2019-0073

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Abstract

ABSTRACTBackground:Neuroblastoma is a pediatric tumor with a mortality rate of 40% in the most aggressive cases. Tumor microenvironment components as immune cells contribute to the tumor progression; thereby, the modulation of immune cells to a pro-inflammatory and antitumoral profile could potentialize the immunotherapy, a suggested approach for high-risk patients. Preview studies showed the antitumoral potential of BJcuL, a C- type lectin isolated from Bothrops jararacussu venom. It was able to induce immunomodulatory responses, promoting the rolling and adhesion of leukocytes and the activation of neutrophils.Methods:SK-N-SH cells were incubated with conditioned media (CM) obtained during the treatment of neutrophils with BJcuL and fMLP, a bacteria-derived peptide highly effective for activating neutrophil functions. Then we evaluated the effect of the same stimulation on the co-cultivation of neutrophils and SK-N-SH cells. Tumor cells were tested for viability, migration, and invasion potential.Results:In the viability assay, only neutrophils treated with BJcuL (24 h) and cultivated with SK-N-SH were cytotoxic. Migration of tumor cells decreased when incubated directly (p < 0.001) or indirectly (p < 0.005) with untreated neutrophils. When invasion potential was evaluated, neutrophils incubated with BJcuL reduced the total number of colonies of SK-N-SH cells following co-cultivation for 24 h (p < 0.005). Treatment with CM resulted in decreased anchorage-free survival following 24 h of treatment (p < 0.001).Conclusion:Data demonstrated that SK-N-SH cells maintain their migratory potential in the face of neutrophil modulation by BJcuL, but their invasive capacity was significantly reduced.

Highlights

Neuroblastoma is a pediatric tumor with a mortality rate of 40% in the most aggressive cases
For both 1 h or 24 h conditions, conditioned media (CM) from neutrophils treated with Bothrops jararacussu lectin (BJcuL) and fMLP improved cell migration compared to CM-RPMI, restoring the migration capacity, closer to RPMI control (Figure 2 and Additional file 1)
Incubation of tumor cells with the conditioned media (CM-RPMI) reduced the number of colonies, which was more pronounced for the CM from neutrophils treated with BJcuL (CM-BJcuL) and fMLP (CMfMLP), with a drop of 40% and 35% in the number of colonies, respectively, when compared with the RPMI control (Figure 4)

Summary

Introduction

Neuroblastoma is a pediatric tumor with a mortality rate of 40% in the most aggressive cases. Preview studies showed the antitumoral potential of BJcuL, a C- type lectin isolated from Bothrops jararacussu venom. BJcuL is a lectin isolated from Bothrops jararacussu snake venom. It is a typical representative of the C-type animal lectin superfamily, having the carbohydrate recognition domain (CRD), and exhibiting specificity for β-d-galactosides [1]. Previous studies have shown that BJcuL has immunomodulatory effects. It can increase the adhesion and rolling of leukocytes in the endothelium of pre-capillary vessels [2], cause edema and increased vascular permeability when injected into mouse hind paw [3]. BJcuL is capable of modulating neutrophils to a pro-inflammatory phenotype, inducing the production of anion superoxide and increased phagocytic function [6]

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